Continuous chromatographic protein refolding.

摘要: Column-based protein refolding requires a continuous processing capability if reasonable quantities of are to be produced. A popular colunm-based method, size-exclusion chromatography (SEC) refolding, employs matrices separate unfolded from denaturant, thus the protein. In this work, we conduct comparison SEC with by batch dilution, using lysozyme as model Lysozyme yield was found extremely sensitive chemical composition buffer and particularly concentration dithiothreitol (DTT) introduced denatured mixture. not adversely affected DTT carry-over small contaminants in solution separated during operation. We also find that, contrary previous reports, on columns leads yields slightly better than dilution at low concentrations but advantage disappears higher concentrations. As batch-mode would limiting step column based downstream process, method translated continuously operating system (preparative annular chromatography, P-CAC). It shown that P-CAC elution profile is similar stationary column, making scale-up translation relatively simple. Moreover, it high (72%) (>1 mg ml(-1)) could obtained. (C) 2003 Elsevier B.V. All rights reserved.